The EFNB1 Knockout A2780 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A2780 human ovarian adenocarcinoma cell line. This product introduces targeted gene disruption of the EFNB1 locus, generating a heterogeneous pool of cells with loss-of-function mutations in ephrin-B1. The polyclonal format reflects a diverse collection of editing outcomes, offering a robust model for studying ephrin-B1 function without clonal bias.
The A2780 cell line is a well-characterized model of high-grade serous ovarian adenocarcinoma, established from a primary tumor of an untreated patient. These cells exhibit epithelial morphology and retain key oncogenic signaling pathways, making them a relevant substrate for investigating ephrin-B1-mediated processes in ovarian cancer biology.
Ephrin-B1 (EFNB1) encodes a transmembrane ligand that engages EphB receptor tyrosine kinases (EphB1?CB3) to initiate bidirectional signaling. Forward signaling through EphB receptors activates SRC and FAK, modulating Rho GTPases (RHOA, RAC1) to regulate actin dynamics and cell adhesion. Reverse signaling via ephrin-B1 recruits adaptor proteins including GIPC1, GRIP1, and SDCBP. EFNB1 expression is transcriptionally controlled by regulators such as HOXA13, MSX2, HIF1A, TGFB1, and WNT signaling. Disruption of EFNB1 abrogates both branches of Eph/ephrin signaling, impairing downstream cascades including MAPK/ERK (MAPK1/3) and PI3K-Akt (AKT), and altering cell adhesion, migration, and proliferation.
In the A2780 ovarian cancer model, EFNB1 knockout disrupts ephrin-B1/EphB-mediated bidirectional signaling, leading to impaired cell adhesion and migration, key processes in metastasis. The loss of ephrin-B1 reduces activation of SRC-FAK and Rho GTPase pathways, attenuating invasive behavior. Additionally, diminished MAPK/ERK and PI3K-Akt signaling may suppress proliferation and survival, potentially reducing tumorigenic capacity. This knockout model therefore enables dissection of ephrin-B1??s role in ovarian cancer progression and the tumor microenvironment.
Researchers can utilize this polyclonal knockout cell population to investigate ephrin-B1-dependent mechanisms in a variety of functional assays. Typical applications include Western blotting to confirm loss of EFNB1 and assess phospho-EphB levels, RT-qPCR to quantify transcript changes in downstream targets such as RHOA, RAC1, and MAPK1/3, and immunofluorescence to visualize alterations in adhesion complex formation. Transwell migration and invasion assays, in combination with phospho-signaling analysis of ERK and Akt, enable functional dissection of EFNB1??s role in motility. Moreover, these cells serve as a platform for screening small molecules targeting EphB receptors or for modeling ephrin-B1-related pathologies like craniofrontonasal syndrome. For further information about assay conditions or lot-specific characteristics, please contact Ascent Research.
